miR-30a-5p alleviating LPS induced acute lung injury by targeting RUNX2
This study aims to investigate the effect of up-regulation of miR-30a-5p on LPS-induced acute lung injury(ALI)and its relationship with human runt related transcription factor 2(RUNX2).For this purpose,lung cell line MRC-5 was transfected with miR-30a-5p mimics and miR-NC,which were recorded as miR-30a-5p group and miR-NC group,respectively.Cells that did not receive any treatment were named blank group.The cells were treated with LPS(1 mg/mL)and divided into LPS+miR-30a-5p group,LPS+miR-NC group,and LPS group.C57BL/6 mice were intraperitoneally injected with LPS to establish the ALI model and subsequently divided into control group,ALI group,ALI+miR-NC group,and ALI+miR-30a-5p group.The lung tissue of mice was H-E stained to examine the pathological changes.The lung injury score was estimated,and the ratio of lung wet weight/dry weight(W/D)was calculated.Serum IL-1β,IL-6,and TNF-α levels were detected by ELISA.The expression levels of miR-30a-5p in cells and tissue were detected by qRT-PCR.The expressions of RUNX2 in tissue and cells were detected by Western blotting.Double luciferase reporter gene test was used to analyze the targeting relationship between miR-30a-5p and RUNX2.The results showed that compared to those of the blank group,the level of miR-30a-5p de-creased and RUNX2 increased in the LPS group(P＜0.01).Compared to those of the miR-NC group and LPS+miR-NC group,the expression of miR-30a-5p increased and RUNX2 decreased in the miR-30a-5p group and LPS+miR-30a-5p group(P＜0.01).Compared to the control group,the ALI group showed decreased miR-30a-5p and increased RUNX2 expression.In addition,the lung injury score,W/D ratio,and IL-6,IL-1β,TNF-α levels all increased in the ALI group(P＜0.01).Compared to those of the ALI+miR-NC group,the level of miR-30a-5p in lung tissue in the ALI+miR-30a-5p group increased,RUNX2 level decreased,and lung injury score,W/D ratio,and TNF-α,IL-1β,IL-6 levels all decreased(P＜0.01).Multiple possible binding sites between miR-30a-5p and RUNX2 3'-UTR were identified.The luciferase activity of the miR-30a-5p+RUNX2-WT group was lower than that of the miR-NC+RUNX2-WT group(P＜0.01).Taken together,miR-30a-5p shows lower expression in the lung tissue of ALI mice and overexpression of miR-30a-5p can improve LPS-induced lung injury in mice by regulating RUNX2.
microRNA-30a-5phuman runt related transcription factor 2acute lung injury
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